• Title/Summary/Keyword: Proteinase

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Antineutrophil Cytoplasmic Antibody in Korean Children with Henoch-$Sch{\ddot{o}}nlein$ Purpura (소아 Henoch-$Sch{\ddot{o}}nlein$ 자반증에서의 항호중구 항체)

  • Choi, Ji-Na;Shin, Jae-Il;Lee, Jae-Seung;Kim, Hyon-Suk
    • Childhood Kidney Diseases
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    • v.12 no.2
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    • pp.164-169
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    • 2008
  • Purpose : There have been some reports on the prevalence of positive antineutrophil cytoplasmic antibody(ANCA) in Henoch-$Sch{\ddot{o}}nlein$ purpura(HSP), but the results were conflicting. We performed this study to evaluate the clinical significance of ANCA(c-ANCA and p-ANCA) in Korean children with HSP. Methods : The medical records of 30 patients(13 boys and 17 girls) aged 6.0$\pm$1.9(5-12) years with a clinical diagnosis of HSP based on the EULAR/PReS criteria were reviewed retrospectively. From the years 2007 to 2008, the sera from children with acute HSP were tested for antineutrophil cytoplasmic antibodies(ANCA). The target antigens of these autoantibodies are proteinase 3(c-ANCA) or myeloperoxidase(p-ANCA). Results : Palpable purpura was seen in all 30 patients(100%), abdominal pain in 20(67%), arthralgia in 17(57%), and renal involvement in 11(37%). Laboratory findings showed leukocytosis in 4 patients(13%), thrombocytosis 18 in(60%), and elevated erythrocyte sedimentation rate in 18(60%). Anti-streptolysin O titers were elevated in 7% of the patients and no patient showed elevation of serum IgA level. The sera from 29 patients were negative for c-ANCA and p-ANCA by indirect immunofluorescence, but only one patient had weakly positive results, which became negative at follow-up. Conclusions : We conclude that c-ANCA or p-ANCA is not an important serologic marker in children with HSP, because it was neither diagnostically nor immunologically specific in children with HSP. These results suggest that ANCA are not involved in the pathogenesis of HSP in children.

A Gelatinase A Isoform, GA110, of Human Follicular Fluid Is Degraded by the Bovine Oviductal Fluid Component (소의 수란관액에 의한 사람 난포액의 Gelatinase A 동위효소인 GA110의 분해)

  • Kim, Min-Jung;Kim, Ji-Young;Leec, Seung-Jae;Yoon, Yong-Dal;Cho, Dong-Jae;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.5 no.1
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    • pp.23-33
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    • 2001
  • When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.

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Loss of the Retinoblastoma Gene in Non-Small Cell Lung Cancer (비소세포폐암에서의 망막모세포종유전자의 소실)

  • Lee, Choon-Taek;Kim, Chang-Min;Zo, Jae-Ill;Shim, Young-Mog;Hong, Weon-Seon;Lee, Jhin-Oh;Kang, Tae-Woong
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.98-103
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    • 1993
  • Background: Inactivation of retinoblastoma gene (Rb) has been observed in a variety of human cancers. Loss of heterozygosity (LOH) of Rb which is a common mode of allelic inactivation of Rb, has been known as a frequent genetic event in small cell lung cancer but it has been detected less frequently in non-small cell lung cancer. To define the role of Rb deletion in lung cancer, we investigated the genomic DNAs of 43 non-small cell lung cancers and 1 small cell lung cancer paired with normal lung tissues obtained by thoracotomy. Methods: The genomic DNAs were obtained by the digestion with proteinase K followed by phenol-chloroform extraction method. The genomic DNAs were digested by restriction endonuclease (EcoRI), separated by agarose gel electrophoresis, transferred to nylon membrane by Southern blot transfer and then hybridized with labelled Rb 1 probe which contains. 1.4 kb sized DNA sequence containing N-terminal portion of Rb. Results: In 26 squamous cell lung cancers, 16 cases were informative after EcoRI digestion and LOH of Rb was found in 10 cases (62.5%). In 17 adenocarcinomas of lung, 11 cases were informative and LOH of Rb was found in five cases (45.4%). The analysis of clinical parameters revealed no significant differences between the two groups with or without LOH of Rb in the aspects of age, sex, degree of differentiation, stage and smoking amount. Conclusions: These results suggest that Rb inactivation is also significantly involved in the molecular pathogenesis of non-small cell lung cancer.

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Mutations of p53 Tumor Suppressor Gene in Human Lung Cancer Cell Lines (사람 폐암세포주에서 p53 종양억제유전자의 변이)

  • Hong, Weon-Seon;Hong, Seok-Il;Lee, Dong-Soon;Son, Young-Sook;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.6
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    • pp.653-658
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    • 1993
  • Background: Recent advancement of molecular genetics has revealed that malignant transformation of a cell may be a complex multistep process and this process is grouped, in general, into two distinct categories, activation of protooncogenes and inactivation of tumor suppressor genes. This study was focused on the mutation of p53 tumor suppressor gene, because p53 gene mutation is now generally accepted to be one of the most frequent genetic changes in a variety of human cancers. Although lung cancer is one of the common cancers in Korea, the genetic change in the carcinogenesis process is not yet known clearly. To investigate the role of p53 gene mutation in lung cancer, we examined the mutations of exon 4-8 of the p53 gene in humna lung cancer cell lines, because most of the mutations of p53 gene have been reported to develop in exon 4-8. Method: Genomic DNA was obtained by the digestion of proteinase K and the extraction by phenol-chloroform-ethanol method from two human pulmonary adenocarcinoma cell lines, PC-9 and PC-14, and one human small cell lung cancer cell line, H69. To detect the mutations of exon 4-8 of the p53 gene, polymerase chain reaction single-strand conformation polymorphism(PCR-SSCP) analysis was performed with the DNA extracted from the cells. Results: The mutation of p53 gene was present in all three cell lines tested. In PC-9, PC-14 and H69, the altered mobility was detected in exon 7, 7 and 5, respectively. Conclusion: These results suggest that p53 gene mutation plays an important role in certain steps of the carcinogenesis of human non-small cell and small cell lung cancer.

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EFFECTS OF SUBINHIBITORY CONCENTRATIONS OF ANTIMICROBIAL AGENTS ON CELL SURFACE PROPERTIES AND VIRULENCE FACTORS OF MUTANS STREPTOCOCCI (아저해농도(亞沮害濃度)의 항균물질이 mutans streptococci의 세포표면성질과 독력인자에 미치는 영향)

  • Kim, Young-Jae;Hahn, Se-Hyun;Lee, Sang-Hoon;Jang, Ki-Taeg;Kim, Chol-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.4
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    • pp.605-616
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    • 2004
  • Subinhibitory concentrations (sub-MICs) refer to concentrations below minimum inhibitory concentrations (MICs). The antimicrobial agents may be present at relatively high concentration, at least higher than bacterial MIC and thereafter be deserted off a surface and function at sub-MICs, perhaps by interfering with bacterial metabolism. Consequently, the aim of this study was to determine the effects of growth, in the presence of sub-MICs of antimicrobial agents, on the cell surface properties and virulence factors of mutans streptococci and to investigate the efficacy of a chemical approach in vitro. Streptococcus mutans Ingbritt and Streptococcus sobrinus 6715-7 were used. Eight antimicrobial agents (Sanguinaria extract;SG, Chlorhexidine digluconate;CHX, Fluoride;F, Propolis;PP, Hydrogen peroxide;HP, Triclosan;TC, Sodium dodecyl sulfate;SDS Cetylpyridinium chloride; CC) were diluted serially in broth to determine MICs and to compare the growth rate, acid production, hydrophobicity, adhesion activity to saliva coated hydroxyapatite, glucan synthesis and cellular aggregation of experiment groups (in the presence of sub-MICs) with those of control (in the absence of antimicrobial agents). Sub-MICs of antimicrobial agents affected the growth of cells, hydrophobicity, and adhesion of bacteria to saliva coated hydroxyapatite and glucan synthesis. They also resulted in a significant reduction in pH after 12 hours (p<0.05). By cells pretreated with proteinase K, either the aggregation induced by antimicrobial agents was completely inhibited or the aggregation titers were markedly increased. According to the results of the present study, each antimicrobial agent at sub-MICs could affect similar as its known action mechanism and could continually inhibit cariogenic bacteria at such concentrations. Thus, the use of these antimicrobial agents would be one of the effective methods to prevent dental caries.

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Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Anti-listeria Activity of Lactococcus lactis Strains Isolated from Kimchi and Characteristics of Partially Purified Bacteriocins (김치에서 분리한 Lactococcus lactis 균주의 항리스테리아 활성 및 부분 정제된 박테리오신의 특성)

  • Son, Na-Yeon;Kim, Tae-Woon;Yuk, Hyun-Gyun
    • Journal of Food Hygiene and Safety
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    • v.37 no.2
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    • pp.97-106
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    • 2022
  • Listeria monocytogenes (L. monocytogenes) is one of gram-positive foodborne pathogens with a very high fatality rate. Unlike most foodborne pathogens, L. monocytogenes is capable of growing at low temperatures, such as in refrigerated foods. Thus, various physical and chemical prevention methods are used in the manufacturing, processing and distribution of food. However, there are limitations to the methods such as possible changes to the food quality and the consumer awareness of synthetic preservatives. Thus, the aim of this study was to evaluate the anti-listeria activity of lactic acid bacteria (LAB) isolated from kimchi and characterize the bacteriocin produced by Lactococcuslactis which is one of isolated strains from kimchi. The analysis on the anti-listeria activity of a total of 36 species (Lactobacillus, Weissella, Lactobacillus, and Lactococcus) isolated from kimchi by the agar overlay method revealed that L. lactis NJ 1-10 and NJ 1-16 had the highest anti-listeria activity. For quantitatively analysis on the anti-listeria activity, NJ 1-10 and NJ 1-16 were co-cultured with L. monocytogenes in Brain Heat Infusion (BHI) broth, respectively. As a result, L. monocytogenes was reduced by 3.0 log CFU/mL in 20 h, lowering the number of bacteria to below the detection limit. Both LAB strains showed anti-listeria activity against 24 serotypes of L. monocytogenes, although the sizes of clear zone was slightly different. No clear zone was observed when the supernatants of both LAB cultures were treated with proteinase-K, indicating that their anti-listerial activities might be due to the production of bacteriocins. Heat stability of the partially purified bacteriocins of NJ 1-10 and NJ 1-16 was relatively stable at 60℃ and 80℃. Yet, their anti-listeria activities were completely lost by 60 min of treatment at 100℃ and 15 min of treatment at 121℃. The analysis on the pH stability showed that their anti-listeria activities were the most stable at pH 4.01, and decreased with the increasing pH value, yet, was not completely lost. Partially purified bacteriocins showed relatively stable anti-listeria activities in acetone, ethanol, and methanol, but their activities were reduced after chloroform treatment, yet was not completely lost. Conclusively, this study revealed that the bacteriocins produced by NJ 1-10 and NJ 1-16 effectively reduced L. monocytogenes, and that they were relatively stable against heat, pH, and organic solvents, therefore implying their potential as a natural antibacterial substance for controlling L. monocytogenes in food.

Effect of Inhibition Macrophage Migration Inhibitory Factor Activation by Hominis Placenta Herbal Acupuncture on Rheumatic Arthritis (자하차약침(紫河車藥鍼)의 MIF 활성 억제를 통한 LPS 유발 류마티스성 관절염의 치료 효과)

  • Hwang, Ji-Hye;Cho, Hyun-Seok;Lee, Hyun-Jin;Lee, Dong-Gun;Jeong, Won-Je;Jung, Chan-Yung;Kim, Kyung-Ho
    • Journal of Acupuncture Research
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    • v.25 no.3
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    • pp.41-51
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    • 2008
  • Objectives : This study is to evaluate Effect of Inhibition Macrophage Migration Inhibitory Factor(MIF) activation by Hominis Placenta Herbal Acupuncture(HPA) on Rheumatic Arthritis(RA). Hominis Placenta is the placenta of healthy human, which is vital-strengthening medical stuff. In recent years, Hominis Placenta applied to chronic diseases because it makes us more resistance to diseases. Therefore it is supposed that HPA is effective on RA, a kind of autoimmune disease. When RA is induced, MIF is activated, too. MIF affects the process of inflammatory disease including RA. Methods : In order to investigate the effect of Hominis Placenta extraction on MIF(early RA inducing cytokine) and MMP(Matrix Metallo Proteinase)-9 mRNA expression by means of Reverse Transcriptase- Polymerase Chain Reaction(RT-PCR). In this study, we investigated the effect of Hominis Placenta extraction on MIF(early RA inducing cytokine) and MMP-9 mRNA expression by means of RT-PCR. Besides we investigated changing of MIF in synovial membrane and, Interleukin-6 receptor(IL-6R)-$\alpha$(pro-inflammatory cytokine), Signal transducers and activators of transcription(STAT)-3, MMP-9 after treating mouse, which is artificially attacked with RA, with HPA on its $ST_{35}$, LE201 in vivo. Results : 1. As a result of treating Lipopolysaccharide(LPS)-stimulated Raw246.7cell with HPA, MIF(RA related cytokine) and MMP-9 mRNA expression is reduced in vitro. And this reaction is concentration-dependatant. 2. In synovial membrane of the mice treated with HPA, inhibition of MIF, IL-6R-$\alpha$, STAT3 & MMP-9 activation is observed in vivo. Conclusions : From the above results, it might be suggested that HPA mitigate tissue damage originated from RA, because it intercepts the early process of by inhibition MIF activity.

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Characterization of a heat-resistant antimicrobial peptide secreted by Bacillus subtilis A405 (Bacillus subtilis A405 균주가 생성하는 내열성 항균 peptide의 특성 검정)

  • Koo, Bon-Sung;Lee, Seung-Bum;Yoon, Sang-Hong;Song, Gae-Kyung;Chung, Dae-Sung;Byun, Myung-Ok;Ryu, Jin-Chang
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.28-35
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    • 1998
  • An antimicrobial peptide producing bacterium, Bacillus subtilis A405, was screened and identified among 700 of antagonistic bacteria. The heat-resistant antimicrobial peptide, AMP-405, was purified from the broth culture of B. subtilis A405 through $20{\sim}40%$ ammonium sulfate precipitation and ultrafiltration. The AMP-405 exhibited strong antimicrobial activities against Botrytis cinerea, Cercospora sp., Fusarium oxysporum, Penicillium digitatum, Celletotrichum gloeosporioides, Rhizoctonia solani, Pythium ultimum, Pyricularia oryzae, Escherichia coli, Pseudomonas spp. and Candida albicans. The molecular weight of the peptide was about 3.0 kDa determined by SDS-PAGE, Native-PAGE and Tris-Tricine gradient electrophoresis, and composed of 9 kinds of amino acid such as aspartic acid, glycine, serine, glutamine, valine, leucine, isoleucine, proline, tyrocine. To determine the efficiency of AMP-405 as a potential maintenance of fruits freshness, cherry tomato was srored at $25^{\circ}C$ for 2 weeks after treatment with $50{\mu}g/ml$ of AMP-405 and $10^{5}$ spores/ml of Botrytis cinerea simultaneously. Treatment with AMP-405 resulted in significantly less infection by Botrytis cinerea, than the treatment with tap water as a control.

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Identification of Genes Differentially Expressed by Oryctes rhinoceros nudivirus Infection in the Korean Rhinoceros Beetle, Allomyrina dichotoma (장수풍뎅이(Korean Rhinoceros Beettle, Allomyrina dichotoma)에 Oryctes rhinoceros nudivirus 감염 특이적으로 발현하는 유전자 동정)

  • Kwon, Kisang;Yoo, Bo-Kyung;Suh, Hyun-Woo;Ko, Young Hwa;Kim, Hong Geun;Lee, Seokhyun;Park, Kwan-Ho;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.25 no.8
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    • pp.942-946
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    • 2015
  • The Korean rhinoceros beetle (Allomyrina dichotoma) is popular as a pet and as a food ingredient, and it is commercially distributed in Korea. It is also traditionally regarded as a medicine for liver-related diseases. Recently, the Oryctes rhinoceros nudivirus was introduced from Southeast Asia. This virus is reported as a disease factor for A. dichotoma in mass-rearing facilities, and economic losses due to this viral infection have been increasing in Korea since the 2010s. In this study, we observed serious structural changes in the fat body and the intestine of virus-infected beetles. We report five genes that are up-regulated by the viral infection in the intestine: BTF3H4-like (transcription factor BTF3 homolog 4-like), SPS-like (serine proteinase stubble-like), COPB1 (coatomer protein complex, subunit beta 1), T-CP (T-complex 1 subunit gamma), and HSP70 HSP70 (heat shock protein 70). The results may provide a clue for the early diagnosis and disease-treatment that occurs in mass-rearing facilities. The improvement of stable productivity will increase the farmers’ income, and quality control of beetle-breeding will help industries to utilize this beetle as a promising food ingredient.